本文選題：人類表皮生長因子2陽性 + 胃癌��； 參考：《浙江大學(xué)》2014年碩士論文

【摘要】：背景： 胃癌是嚴重危害人類生命健康的重大疾病,世界衛(wèi)生組織癌控項目數(shù)據(jù)顯示,全球每年胃癌新發(fā)病例約93萬,死亡人數(shù)約70萬,占所有癌癥死亡率的1/10;我國每年新發(fā)胃癌患者達40萬人,死亡人數(shù)達30萬,患病率和死亡率均是世界平均水平的2倍多。目前在我國胃癌早期檢出率不到10%。超過90%以上的胃癌患者于就診時已是進展期以上,或在確診時已經(jīng)發(fā)生了轉(zhuǎn)移或播散,從而失去了手術(shù)的機會。因此,胃癌的化學(xué)治療是國內(nèi)絕大多數(shù)胃癌患者的主要治療方法,在我國有著重要的研究意義。但是,當前臨床應(yīng)用的常規(guī)化療藥物全身毒副作用較大,患者耐受性差；腫瘤細胞對化療藥物產(chǎn)生耐藥等,都容易導(dǎo)致化療的失敗。針對胃癌常規(guī)化療藥物的全身用量大,毒副作用大的研究,具有極大地學(xué)術(shù)價值、社會效應(yīng)及應(yīng)用前景。近年來,由于各種納米粒子能理想地在體內(nèi)傳遞各種物質(zhì)并具有可調(diào)的表面化學(xué)靶向結(jié)構(gòu),使得它在腫瘤治療中凸顯出獨特的優(yōu)勢。 目的： 本研究在胃癌細胞人類表皮生長因子2受體為靶向的基礎(chǔ)上,以貴金屬Au納米顆粒通過交換偶合與Fe304形成納米復(fù)合物作為載體,化學(xué)結(jié)合單克隆抗體赫賽汀及一線化療藥物奧沙利鉑,從而構(gòu)建以人類表皮生長因子2為靶向的磁共振成像示蹤控釋奧沙利鉑的新型納米復(fù)合物Oxaliplatin-Au-Fe3O4-Herceptin,并且評價其在人類表皮生長因子2陽性胃癌中的價值。 方法： 1.應(yīng)用交換偶合法及熱降解技術(shù)在金表面降解乙酰丙酮鐵完成Au-Fe3O4金屬納米復(fù)合物的構(gòu)建。通過表面修飾劑的加入對Au-Fe3O4納米顆粒的形貌進行調(diào)控。透射電鏡表征納米粒子。 2.液相Oxaliplatin-Au-Fe3O4-Herceptin與1-(3-二甲氨基丙基)-3-乙基二亞胺及N-羥基硫代琥珀�；旌闲纬蛇B接鍵,再與赫賽汀溶液混合培養(yǎng),形成Au-Fe3O4-Herceptin�？焖賹游龇兓玫綂W沙利鉑連接配體�；旌蠆W沙利鉑混懸液、Au-Fe3O4-Herceptin及連接配體,優(yōu)化比例,培養(yǎng)純化得到Oxaliplatin-Au-Fe3O4-Herceptin。MALDI-MS驗證產(chǎn)物,體外透析袋不同pH值下檢測奧沙利鉑結(jié)合效率。 3. MTS法分別驗證Au-Fe3O4、Au-Fe3O4-Herceptin Oxaliplatin-Au-Fe3O4-Herceptin細胞毒性。 4.常規(guī)培養(yǎng)HER2陽性胃癌細胞系SGC-7901。分別與Au-Fe3O4、 Oxaliplatin-Au-Fe3O4-Herceptin孵育2小時,透射電鏡觀察納米復(fù)合物在胃癌細胞表面胞吞作用的程度。 5.胃癌原位移植瘤模型的建立及分組：研究不同時限注射Oxaliplatin-Au-Fe3O4-Herceptin的胃癌裸鼠模型,調(diào)整建立磁共振掃描參數(shù),觀察小鼠腫瘤組織變化。結(jié)果 1.透射電鏡下,見納米粒子大小約25nm,鐵粒子與金粒子1:1恒定； 2.成功構(gòu)建了Oxaliplatin-Au-Fe3O4-Herceptin納米復(fù)合物,并通過質(zhì)譜、紅外及透析袋緩釋奧沙利鉑證實了各分子間通過化學(xué)連接鍵相連。細胞毒性實驗顯示：當鐵離子濃度在30ug/ml以內(nèi)時,Oxaliplatin-Au-Fe3O4-Herceptin納米復(fù)合物對細胞基本沒有毒性； 3.體外胞吞實驗：將Oxaliplatin-Au-Fe3O4-Herceptin納米復(fù)合物、Au-Fe3O4納米粒子以相同鐵粒子濃度與SGC-7901細胞共孵育2h后透射電鏡可見,Oxaliplatin-Au-Fe3O4-Herceptin納米復(fù)合物被大量吞入細胞,而Au-Fe3O4納米粒子未見被吞噬入細胞內(nèi)； 4.小鼠尾靜脈注射納米粒子復(fù)合物一段時間后,磁共振掃描老鼠皮下移植瘤,見腫瘤組織內(nèi)T2相低密度信號灶。腫瘤組織普魯士染色后可見腫瘤血管旁大量鐵粒子聚集。結(jié)論： 本課題通過學(xué)科交叉研究,研制成功針對人類表皮生長因子2受體金屬納米化合物Oxaliplatin-Au-Fe3O4-Herceptin,能結(jié)合奧沙利鉑靶向胃癌,并實現(xiàn)磁共振示蹤控釋,獲得擁有自主知識產(chǎn)權(quán)。研究明確了該種化合物的體內(nèi)外作用作用機制和生物安全性,為進一步的臨床應(yīng)用和人類表皮生長因子2陽性胃癌患者的個性化治療,以及新藥的開發(fā)提供堅實的實驗基礎(chǔ)。同時,亦為開發(fā)新型靶向治療藥物和靶向治療提供新的思路和技術(shù)支持。
[Abstract]:Background :

Gastric cancer is a major disease which seriously endangers the health of human life . The data of World Health Organization Cancer Control Project shows that the number of new cases of gastric cancer in the world is about 9.3 million , the number of deaths is about 700,000 , accounting for 1 / 10 of all cancer mortality . In our country , more than 90 % of patients with gastric cancer have progressed more than 10 % each year .
In recent years , various kinds of nano - particles can transfer various substances in vivo and have adjustable surface chemical targeting structure , so that it has unique advantages in tumor treatment .

Purpose :

Based on the targeting of human epidermal growth factor - 2 receptor in gastric cancer cells , a novel nano - complex Oxhide - Au - Fe _ 3O _ ( 4 - -Fe _ 3O _ 3O _ 4 - ) was prepared by exchange coupling with Fe304 as a carrier , and it was used as a carrier to construct a novel nano - composite Oxhide - Au - Fe3O4 - Hercee with magnetic resonance imaging and release oxaliplatin targeting human epidermal growth factor 2 , and its value in human epidermal growth factor 2 positive gastric cancer was evaluated .

Method :

1 . The Au - Fe _ 3O _ 4 metal nanoparticles were prepared by using exchange coupling method and thermal degradation technique on gold surface . The morphology of Au - Fe3O4 nanoparticles was controlled by the addition of surface modifier . The nano - particles were characterized by TEM .

2 . The preparation of Oxhide - Au - Fe _ 3O _ 3 - ethyldiimine with 1 - ( 3 - dimethylaminopropyl ) -3 - ethyldiimine and N - hydroxythiosuccinyl was carried out to form the bond , and then mixed with Hercystatin solution to form Au - Fe _ 3O _ 3O _ 4 - HerceII . Rapid chromatographic method was used to purify the oxaliplatin - linked ligand .

3 . The toxicity of Au - Fe _ 3O _ 4 , Au - Fe _ 3O _ 3O _ 3O _ 3 - 2 - Fe _ 3O _ 3O _ 3O _ 3 - 4 - Fe _ 3O _ 3O _ 4 - 2 was verified by MTS method .

4 . The HER2 - positive gastric cancer cell line SGC - 790was incubated with Au - Fe _ 3O _ 3O _ 4 and Oxhide - Au - Fe _ 3O _ 3O _ 3O _ 3O _ 4 for 2 hours , and the extent of endocytosis of the nano - complexes on the surface of gastric cancer cells was observed by transmission electron microscope .

5 . Establishment and grouping of in - situ transplanted tumor model of gastric cancer : The nude mouse model of gastric cancer with different time - frame injection of Oxhide - Au - Fe3O4 - HerceII was studied , and the parameters of magnetic resonance scan were adjusted to observe the changes of tumor tissues in mice .

1 . Under the transmission electron microscope , the size of the nanoparticles is about 25nm , and the iron particles are constant with the gold particles 1 : 1 ;


2 . Oxhide - Au - Fe _ 3O _ 4 - Hertin nanocomposites were successfully constructed . Through mass spectrometry , IR and slow release of oxaliplatin , it was confirmed that there was no toxicity to the cells .


3 . In vitro endocytosis experiment : Oxhide - Au - Fe _ 3O _ 3O _ 4 - Hertin nanocomposites were incubated for 2 hours at the same concentration of Fe particles and SGC - 790cells for 2 hours . The Oxhide - Au - Fe _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 4 nanoparticles were absorbed into the cells and the Au - Fe _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 3O _ 4 nanoparticles were swallowed into the cells .


4 . After intravenous injection of nano - particle complex for a period of time , magnetic resonance scanning the mouse subcutaneous transplantation tumor to see the T2 - phase low - density signal focus in the tumor tissue . After the tumor tissue , the tumor tissue was stained and the large amount of iron particles near the tumor vessel were collected . Conclusion :

The study on the mechanism and biological safety of human epidermal growth factor - 2 receptor metal nano - compound Oxhide - Au - Fe _ 3O _ 3O _ 3O _ 4 - Her2 combined with oxaliplatin targeted gastric cancer , and achieved independent intellectual property rights . It also provides a solid experimental basis for further clinical application and personalized treatment of human epidermal growth factor 2 positive gastric cancer patients , and provides new ideas and technical support for the development of new targeted therapy drugs and targeted therapy .
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：TB383.1

【共引文獻】

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2 劉雄雄;李強;;氧化應(yīng)激引起的遺傳變化和DNA甲基化改變在腫瘤發(fā)生中的作用[J];輻射研究與輻射工藝學(xué)報;2013年06期

3 張亮;杜佳;周立為;程q，

本文編號：1989494